Department of Epidemiology, Geisel School of Medicine at Dartmouth College, Lebanon New Hampshire.Department of Pharmacology and Toxicology, Geisel School of Medicine at Dartmouth College, New Hampshire.Department of Community and Family Medicine, Geisel School of Medicine at Dartmouth College, Lebanon New Hampshire.

Department of Epidemiology, Geisel School of Medicine at Dartmouth College, Lebanon New Hampshire.Department of Pharmacology and Toxicology, Geisel School of Medicine at Dartmouth College, New Hampshire.Department of Community and Family Medicine, Geisel School of Medicine at Dartmouth College, Lebanon New Hampshire.

Department of Epidemiology, Geisel School of Medicine at Dartmouth College, Lebanon New Hampshire.Department of Community and Family Medicine, Geisel School of Medicine at Dartmouth College, Lebanon New Hampshire.

Article Figures & Data

Figures

Conceptual diagram illustrating the various steps involved in estimating the mdNLR. In Step 1, putative leukocyte differentially methylated regions (L-DMR) are identified between monocytes, granulocytes, and lymphocytes. Step 2 involves computing the within cell type mean methylation beta-values for each of the putative L-DMRs identified in Step 1. In Step 3, the within cell type mean methylation beta-values are used in conjunction with Houseman and colleagues (23) to predict the proportion monocytes, granulocytes, and lymphocytes for a sample consisting of DNA methylation signatures profiled in WB. In Step 4, the mdNLR is calculated as the ratio of the predicted proportions of granulocytes and lymphocytes and finally, in Step 5, the estimated mdNLR is examined with respect to its association with cancer risk, outcomes, or other clinical variables of interest.

Results obtained from examining the association between mdNLR and HNSCC case–control status and survival. A, Distribution of the predicted proportion of lymphocytes, monocytes, and granulocytes between HNSCC cases and age-matched cancer-free controls. B, Distribution of the mdNLR between HNSCC cases and age-matched cancer-free controls. C, ROC curves demonstrating the ability of mdNLR to correctly classify HNSCC cases from cancer-free controls. Each ROC curve was generated from a different classifier: red ROC curve was based on a classifier that used covariates only (i.e., age, gender, smoking history, and HPV16 status), black ROC curve was from a classifier based on mdNLR only, and green curve was based on a classifier using both mdNLR and the aforementioned covariates. D, Scatter plot of the –log10 log-rank P value as a function of the mdNLR breakpoint used to determine mdNLR low and high groups. Blue line represents the estimated Lowess smoothed curve. E, Kaplan–Meier survival curves for the HNSCC cases in the mdNLR high and low groups.

Results obtained from examining the association between mdNLR and bladder cancer case–control status and survival. A, Distribution of the predicted proportion of lymphocytes, monocytes, and granulocytes between bladder cancer cases and cancer-free controls. B, Distribution of the mdNLR between bladder cancer cases and cancer-free controls. C, Distribution of the mdNLR among bladder cancer cases that died during the follow-up period compared with those that survived or were censored. D, Scatter plot of the –log10 log-rank P value as a function of the mdNLR breakpoint used to determine mdNLR low and high groups. Blue line represents the estimated Lowess smoothed curve. E, Kaplan–Meier survival curves for bladder cancer cases in the mdNLR high and low groups.

Results from the analysis of the mdNLR within the ovarian cancer, breast cancer, and aging data sets. A, Distribution of the mdNLR between pretreatment ovarian cancer cases, posttreatment ovarian cancer cases, and age-matched cancer-free controls. B, ROC curves demonstrating the ability of mdNLR to correctly classify controls versus pretreatment ovarian cancer cases (red line), controls versus posttreatment ovarian cancer cases (black line), and pre- versus posttreatment ovarian cancer cases (green line). C, Scatter plot of the within twin-pair difference in the mdNLR between breast cancer cases and controls (y-axis) as a function of WB sample collection relative to cancer diagnosis (x-axis). Green quadrant indicates that samples were collected prediagnosis and that the mdNLR was higher among the twin-pair member that would eventually be diagnosed with breast cancer. Red quadrant indicates that samples were collected prediagnosis and that the mdNLR was lower among the twin-pair member that would eventually be diagnosed with breast cancer. Blue quadrant indicates that samples were collected postdiagnosis and that the mdNLR was higher among the twin-pair member diagnosed with breast cancer. Purple quadrant indicates that samples were collected postdiagnosis and that the mdNLR was lower among the twin-pair member diagnosed with breast cancer. Blue line represents the Lowess smoothed curve estimated using all 13 twin pairs. D, Distribution of mdNLR as a function of age group among the samples in the aging study. E, Distribution of mdNLR as a function ethnic background among the samples in the aging study.